PKD1基因与抑癌基因P53多态性与非综合征性唇腭裂的关联研究

目的:探讨多囊性肾病基因1(polystic kidney disease,PKD1)多态位点rs8049367与抑癌基因P53(anti-oncogene P53)多态位点rs4791774单核苷酸多态性与中国北方人群非综合征性唇腭裂(NSCL/P)的相关性。方法:运用聚合酶链式反应-连接酶检测反应(PCR-LDR)的检测方法,对602例NSCL/P患者和510例对照人群的PKD1基因的rs8049367位点和抑癌基因P53的rs4791774位点进行基因分型。利用SPSS12.0软件分析PKD1基因,抑癌基因P53多态性与NSCL/P的相关性。结果:rs8049367位点和rs4791774位点基因型及基因频率在两组的分布中差异无统计学意义(P0.05)。结论:PKD1基因的rs8049367位点和抑癌基因P53的rs4791774位点单核苷酸多态性可能与中国北方人群非综合征性唇腭裂的发生无相关性。     

双侧股神经阻滞用于双膝关节置换术患者的麻醉效果和对血清炎性因子水平的影响

目的:研究双侧股神经阻滞术用于双膝关节置换术患者麻醉效果和对患者血清炎性因子水平的影响。方法:选择2015年10月～2018年10月在我院进行双膝关节置换术的110例患者,按照其入院顺序经随机数字表法分为两组,每组55例。对照组采用全身麻醉,研究组采用双侧股神经阻滞联合全身麻醉。比较两组的麻醉情况,治疗前后血清炎性因子白介素6(IL-6)、C反应蛋白(CRP)、舒张压(DBP)、收缩压(SBP)、心率(HR)水平的变化。结果:两组麻醉时间比较差异无统计学意义(P0.05);研究组拔管、恢复室停留和苏醒时间均显著短于对照组(P0.05)。两组术后24 h、48 h血清炎性因子IL-6、CRP水平均高于术前,但研究组以上指标均显著低于对照组(P0.05);两组术中DBP、SBP、HR水平均较术前显著降低(P0.05),但研究组DBP、SBP、HR水平均显著高于对照组(P0.05),两组术后DBP、SBP、HR水平比较差异均无统计学意义(P0.05)。结论:与单纯采用全身麻醉相比,双侧股神经阻滞可有效改善双膝关节置换术患者的麻醉效果,并降低其血清炎症因子和稳定其血流动力学。     

Effects of low-level combined static and weak low-frequency alternating magnetic fields on cytokine production and tumor development in mice

We investigated the effects of weak combined magnetic fields (MFs) produced by superimposing a constant MF (in the range 30 - 150 µT) and an alternating MF (100 or 200 nT) on cytokine production in healthy Balb/C male mice exposed 2 h daily for 14 days. The alternating magnetic field was a sum of several frequencies (ranging from 2.5 - 17.5 Hz). The frequencies of the alternating magnetic field were calculated formally based on the cyclotron resonance of ions of free amino acids (glutamic and aspartic acids, arginine, lysine, histidine, and tyrosine). The selection of different intensity and frequency combinations of constant and alternating magnetic fields was performed to find the optimal characteristics for cytokine production stimulation in immune cells. MF with a constant component of 60 μT and an alternating component of 100 nT, which was a sum of six frequencies (from 5 to 7 Hz), was found to stimulate the production of tumor necrosis factor-α, interferon-gamma, interleukin-2, and interleukin-3 in healthy mouse cells and induce cytokine accumulation in blood plasma. Then, we studied the effect of this MF on tumor-bearing mice with solid tumors induced by Ehrlich ascite carcinoma cells by observing tumor development processes, including tumor size, mouse survival rate, and average lifespan. Tumor-bearing mice exposed to a combined constant magnetic field of 60 μT and an alternating magnetic field of 100 nT containing six frequencies showed a strong suppression of tumor growth with an increase in survival rate and enhancement of average lifespan.     

High-throughput Identification of FLT3 Wild-type and Mutant Kinase Substrate Preferences and Application to Design of Sensitive In Vitro Kinase Assay Substrates

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Highlights

• •Developed a data processing pipeline to format phosphopeptide identifications.
• •Identified the preferred substrate motif for FLT3 and mutant kinases.
• •Designed and validated a panel of pan-FTL3 artificial substrates.
• •Monitored FLT3 and mutant kinase activity through FAStide phosphorylation.

     

Proteasome Inhibitor PS-341 Effectively Blocks Infection by the Severe Fever with Thrombocytopenia Syndrome Virus

Severe fever with thrombocytopenia syndrome(SFTS) is an emerging hemorrhagic fever disease caused by SFTSV, a newly discovered phlebovirus that is named after the disease. Currently, no effective vaccines or drugs are available for use against SFTSV infection, as our understanding of the viral pathogenesis is limited. Bortezomib(PS-341), a dipeptideboronic acid analog, is the first clinically approved proteasome inhibitor for use in humans. In this study, the antiviral efficacy of PS-341 against SFTSV infection was tested in human embryonic kidney HEK293 T(293 T) cells. We employed four different assays to analyze the antiviral ability of PS-341 and determined that PS-341 inhibited the proliferation of SFTSV in 293 T cells under various treatment conditions. Although PS-341 did not affect the virus absorption, PS-341 treatment within a non-toxic concentration range resulted in a significant reduction of progeny viral titers in infected cells.Dual-luciferase reporter assays and Western blot analysis revealed that PS-341 could reverse the SFTSV-encoded nonstructural protein(NS) mediated degradation of retinoic acid-inducible gene-1(RIG-I), thereby antagonizing the inhibitory effect of NSs on interferons and blocking virus replication. In addition, we observed that inhibition of apoptosis promotes virus replication. These results indicate that targeting of cellular interferon pathways and apoptosis during acute infection might serve as the bases of future therapeutics for the treatment of SFTSV infections.     

基于质谱的蛋白质N末端乙酰化程度定量方法的研究

随着质谱技术及各种定量方法的不断完善和发展,定量蛋白质组学的方法不断地被应用到各类生物学研究中。蛋白质组学定性定量数据的处理主要通过一些多功能的商业化或者开源软件来进行,如常用的数据分析软件Proteome Discoverer和Maxquant。但是在通过化学标记对蛋白质N末端乙酰化程度进行定量这一方面,Proteome Discoverer和Maxquant在一定程度上存在准确性不高和完整度不够的问题。于是本研究针对自己的实验特点,通过Java算法编写了相应的定量程序Acequant来完成N末端乙酰化程度的相对定量。本研究将该程序在已有相关报道的He La cell上进行了验证,Acequant共定量到1 587个蛋白质N末端,而Proteome Discoverer和Maxquant分别只定量到42个和306个N末端。同时,手动验证原始图谱也证实了Acequant定量的准确性更好。于是,本研究将此方法进一步应用到秀丽隐杆线虫N末端乙酰化的研究中,并初步发现了线虫整体的N末端乙酰化状态,为进一步的N末端研究提供了支持。     

New insights into the binding mode of pyridine-3-carboxamide inhibitors of E. coli DNA gyrase

Previously we have reported on a series of pyridine-3-carboxamide inhibitors of DNA gyrase and DNA topoisomerase IV that were designed using a computational de novo design approach and which showed promising antibacterial properties. Herein we describe the synthesis of additional examples from this series aimed specifically at DNA gyrase, along with crystal structures confirming the predicted mode of binding and in vitro ADME data which describe the drug-likeness of these compounds.     

ELK-1/JNK/c-Fos对左归降糖解郁方(ZGJTJYF)体外培养的模拟糖尿病并发抑郁症(DD)大鼠海马神经元凋亡中的作用

目的:研究信号通路ELK-1/JNK/c-Fos在左归降糖解郁方(ZGJTJYF)对模拟糖尿病并发抑郁症(DD)环境下抗海马神经元凋亡中的作用。方法:原代培养海马神经元,加入高糖(150 mmol/L)+皮质酮(200μmol/L),构建DD体外细胞模型;将培养的海马神经元细胞随机分为5组:空白血清组、正常组、左归降糖解郁方含药血清组、阳性药(二甲双胍+氟西汀)含药血清组和模型组(每组3个复孔)。模型组和正常组给予等量培养液,其余组加入相应体积分数10%的相应血清,均干预18 h。分别采用Hoechst染色、高内涵细胞成像分析技术和RT-PCR技术分别检测海马神经元凋亡情况及检测凋亡相关ELK-1、JNK和c-Fos蛋白和基因的表达。结果:与空白组比较,模型组细胞凋亡明显,其海马神经元凋亡数量明显增多,ELK-1、JNK和c-Fos的mRNA及蛋白表达水平均显著升高(P<0.05);与模型组比较,左归降糖解郁方含药血清组和阳性药含药血清组大鼠海马神经元可见局部亮点明显减少,凋亡细胞数显著减少,ELK-1、JNK和c-Fos蛋白及mRNA表达均明显下调(P<0.05),且神经网络及树突连接情况得到明显改善。结论:左归降糖解郁方可以减低DD环境下海马神经元中Elk-1、JNK、c-fos表达而起到抗凋亡的效果。     

茯神挥发性成分及其生物活性研究

为研究茯神挥发性成分含量、构成及其生物活性,本实验采用二氧化碳超临界的方法提取茯神低极性成分,以提取率作为响应值,在单因素试验的基础上采用响应曲面法考察提取压力、温度、二氧化碳流速对提取效果的影响,优化提取工艺。运用气相色谱-质谱联用仪(GC-MS)对其主要化学成分进行鉴定,并采用生长速率法测定了挥发油对5种真菌的生物活性。得到提取模型极值点,即提取压力27.26MPa、提取温度55.97℃、提取流速10.68L/min时,提取率达到最大,提取率预测值为1.66mg/g。通过NIST14质谱库检索,鉴定了其中17个主要化合物,运用峰面积归一法确定各个组分的含量,占挥发油总量的84.5%。抗菌实验表明茯神挥发油对采绒革盖菌菌株有低浓度促进高浓度抑制的活性。